vanillate

• 分子结构分类: 有机化合物 - 苯类化合物 - 苯和取代衍生物
• 英文名称: vanillate
• 英文别名: 4-carboxy-2-methoxyphenolate
• 化学式: C₈H₇O₄
• 分子量: 167.141
• InChiKey: WKOLLVMJNQIZCI-UHFFFAOYSA-M

计算性质

• 辛醇/水分配系数(LogP)：
  2.1
• 重原子数：
  12
• 可旋转键数：
  2
• 环数：
  1.0
• sp3杂化的碳原子比例：
  0.12
• 拓扑面积：
  69.6
• 氢给体数：
  1
• 氢受体数：
  4

反应信息

• 作为反应物：
  描述：

  vanillate 在 还原型辅酶II(NADPH)四钠盐 作用下， 反应 48.0h， 生成 4-羟基-3-甲氧基苄醇
  参考文献：
  名称：

  大肠杆菌工程菌株中芳香醛的合成和积累

  摘要：

  芳香醛可用于多种应用，尤其是用作调味剂、香料和药物前体。然而，醛类的微生物合成受到醛类向其相应醇类的快速、内源性和冗余转化的阻碍。我们报告了具有减少的芳香醛还原 (RARE) 的大肠杆菌 K-12 MG1655 菌株的构建，该菌株可作为芳香醛生物合成的平台。对模型底物苯甲醛具有报告活性的六个基因被合理地靶向缺失：三个编码醛酮还原酶的基因和三个编码醇脱氢酶的基因。在 RARE 菌株中表达重组羧酸还原酶并在生长过程中加入苯甲酸盐后，24 小时后苯甲醛仍留在培养物中，苯甲醛转化为苯甲醇的转化率低于 12%。尽管个体过表达结果表明所有六个基因都有助于体内苯甲醛的减少，但以子集缺失菌株为特征的其他实验表明，在测试条件下，两个基因缺失是可有可无的。接下来研究了工程菌株从香草酸中生产香草醛的情况，并成功地防止了副产物香草醇的形成。引入了直接从葡萄糖生物合成香草醛的途径，当使用 RARE 菌株与野生型菌株相比时，香草醛效价提高了

  DOI：

  10.1021/ja506664a
• 作为产物：
  描述：

  5-carboxyvanillate 在 recombinant 5-carboxyvanillate decarboxylase from Sphingomonas paucimobilis SYK-6 、 manganese(ll) chloride 作用下， 生成 vanillate
  参考文献：
  名称：

  5-羧基香草酸脱羧酶的底物畸变及催化反应机制

  摘要：

  5-羧基香草酸脱羧酶 (LigW) 在木质素降解的生化途径中催化 5-羧基香草酸转化为香草酸。该酶被证明需要 Mn2+ 才能发挥催化活性，并且需要少动鞘氨醇单胞菌 SYK-6 酶脱羧的动力学常数（kcat = 2.2 s–1 和 kcat/Km = 4.0 × 104 M–1 s– 1) 和 Novosphingobiumaromaticivorans (kcat = 27 s–1 和 kcat/Km = 1.1 × 105 M–1 s–1) 被确定。在活性位点是否存在结合配体的情况下测定了两种酶的三维结构。来自 N.aromaticivorans 的 LigW 结构与底物类似物 5-硝基香草酸盐 (Kd = 5.0 nM) 结合，测定分辨率为 1.07 Å。该复合物的结构显示出显着的酶诱导硝基取代基从苯环平面变形约 23°。基于在活性位点结合配体存在下确定的高分辨率 X 射线结构、活性位

  DOI：

  10.1021/jacs.5b08251

文献信息

• Identification of UGT84A13 as a candidate enzyme for the first committed step of gallotannin biosynthesis in pedunculate oak (Quercus robur)
  作者：Juliane Mittasch、Christoph Böttcher、Nadezhda Frolova、Markus Bönn、Carsten Milkowski

  DOI：10.1016/j.phytochem.2013.11.023

  日期：2014.3

  UDP-glucose:gallic acid glucosyltransferase activity. UGT84A13 was functionally expressed in Escherichia coli as N-terminal His-tagged protein. In vitro kinetic measurements with the purified recombinant enzyme revealed a clear preference for hydroxybenzoic acids as glucosyl acceptor in comparison to hydroxycinnamic acids. Of the preferred in vitro substrates, protocatechuic, vanillic and gallic acid, only

  编码形成酯的羟基苯甲酸葡萄糖基转移酶 UGT84A13 的 cDNA 是从 Quercus robur 膨胀芽和幼叶的 cDNA 文库中分离出来的。该酶与来自葡萄属的白藜芦醇/羟基肉桂酸酯和羟基苯甲酸酯/羟基肉桂酸酯葡萄糖基转移酶显示出高度的序列同一性，并聚集在植物葡萄糖基转移酶的系统发育组 L 中，主要参与 1-O-β-D-葡萄糖酯的形成。计算机转录组分析证实了 UGT84A13 在 Quercus 组织中的表达，该组织之前显示出 UDP-葡萄糖：没食子酸葡萄糖基转移酶活性。UGT84A13 在大肠杆菌中作为 N 端 His 标记蛋白在功能上表达。用纯化的重组酶进行的体外动力学测量显示，与羟基肉桂酸相比，羟基苯甲酸明显更倾向于作为葡萄糖基受体。在优选的体外底物原儿茶酸、香草酸和没食子酸中，只有后者及其相应的 1-O-β-D-葡萄糖酯被发现在年轻的橡树叶中积累。这表明在植物中 UGT84A13
• Salivary Aldehyde Dehydrogenase: Activity towards Aromatic Aldehydes and Comparison with Recombinant ALDH3A1
  作者：Joanna Giebułtowicz、Renata Wolinowska、Anna Sztybor、Monika Pietrzak、Piotr Wroczyński、Jacek Wierzchowski

  DOI：10.3390/molecules14072363

  日期：——

  A series of aromatic aldehydes was examined as substrates for salivary aldehyde dehydrogenase (sALDH) and the recombinant ALDH3A1. Para-substituted benzaldehydes, cinnamic aldehyde and 2-naphthaldehydes were found to be excellent substrates, and kinetic parameters for both salivary and recombinant ALDH were nearly identical. It was demonstrated that for the fluorogenic naphthaldehydes the only produced

  检查了一系列芳香醛作为唾液醛脱氢酶 (sALDH) 和重组 ALDH3A1 的底物。发现对位取代的苯甲醛、肉桂醛和 2-萘醛是极好的底物，唾液和重组 ALDH 的动力学参数几乎相同。已经证明，对于荧光性萘醛，在唾液中孵育后唯一产生的反应产物是羧酸盐。
• Cloning and sequencing of Pseudomonas genes encoding vanillate demethylase
  作者：F Brunel、J Davison

  DOI：10.1128/jb.170.10.4924-4930.1988

  日期：1988.10

  A 2,598-base-pair (bp) SalI-HincII DNA fragment has been cloned which codes for vanillate demethylase, the enzyme responsible for the demethylation of vanillate (3-methoxy-4-hydroxybenzoate) to protocatechuate (3,4-dihydroxybenzoate). Complementation and insertional inactivation experiments have shown that this fragment carries two genes (vanA and vanB) which are predominantly cotranscribed from a promoter upstream of vanA. Nucleotide sequencing of the SalI-HincII fragment confirmed the genetic data: two open reading frames of 987 and 942 bp were present in the transcribed orientation. These had a very high G + C content in the third base of each codon, which is characteristic of Pseudomonas chromosomal genes. Expression of the genes in Escherichia coli with the T7 RNA polymerase-promoter system gave rise to two polypeptides of 36 and 33 kilodaltons which could be identified by deletion analysis as the products of vanA and vanB, respectively. A search of the protein sequence data bank indicated that the vanB gene product was related to the ferredoxin family.

  已经克隆了一个长度为2,598个碱基对的SalI-HincII DNA片段，其中编码香草酸去甲基酶，这种酶负责将香草酸（3-甲氧基-4-羟基苯甲酸）去甲基成原儿茶酸（3,4-二羟基苯甲酸）。补全和插入失活实验表明，该片段携带两个基因（vanA和vanB），这些基因主要从upstream的vanA启动子共同转录。SalI-HincII片段的核苷酸测序证实了基因数据：两个开放阅读框（ORF）分别为987和942个碱基对，并且处于转录方向。这些具有非常高的G+C含量，这是假单胞菌染色体基因的特征。使用T7 RNA聚合酶-启动子系统在大肠杆菌中表达这些基因，产生了两种多肽，分别为36和33千道尔顿，通过缺失分析确定为vanA和vanB的产物。蛋白质序列数据库的搜索表明，vanB基因产物与ferredoxin家族有关。
• Molecular characterization of genes of Pseudomonas sp. strain HR199 involved in bioconversion of vanillin to protocatechuate
  作者：H Priefert、J Rabenhorst、A Steinbüchel

  DOI：10.1128/jb.179.8.2595-2607.1997

  日期：1997.4

  The gene loci vdh, vanA, and vanB, which are involved in the bioconversion of vanillin to protocatechuate by Pseudomonas sp. strain HR199 (DSM 7063), were identified as the structural genes of a novel vanillin dehydrogenase (vdh) and the two subunits of a vanillate demethylase (vanA and vanB), respectively. These genes were localized on an EcoRI fragment (E230), which was cloned from a Pseudomonas sp. strain HR199 genomic library in the cosmid pVK100. The vdh gene was identified on a subfragment (HE35) of E230, and the vanA and vanB genes were localized on a different subfragment (H110) of E230. The nucleotide sequences of fragment HE35 and part of fragment H110 were determined, revealing open reading frames of 1062, 951, and 1446 bp, representing vanA, vanB, and vdh, respectively. The vdh gene was organized in one operon together with a fourth open reading frame (ORF2), of 735 bp, which was located upstream of vdh. The deduced amino acid sequences of vanA and vanB exhibited 78.8 and 62.1% amino acid identity, respectively, to the corresponding gene products from Pseudomonas sp. strain ATCC 19151 (F. Brunel and J. Davison, J. Bacteriol. 170:4924-4930, 1988). The deduced amino acid sequence of the vdh gene exhibited up to 35.3% amino acid identity to aldehyde dehydrogenases from different sources. The deduced amino acid sequence of ORF2 exhibited up to 28.4% amino acid identity to those of enoyl coenzyme A hydratases. Escherichia coli strains harboring fragment E230 cloned in pBluescript SK- converted vanillin to protocatechuate via vanillate, indicating the functional expression of vdh, vanA, and vanB in E. coli. High expression of vdh in E. coli was achieved with HE35 cloned in pBluescript SK-. The resulting recombinant strains converted vanillin to vanillate at a rate of up to 0.3 micromol per min per ml of culture. Transfer of vanA, vanB, and vdh to Alcaligenes eutrophus and to different Pseudomonas strains, which were unable to utilize vanillin or vanillate as carbon sources, respectively, conferred the ability to grow on these substrates to these bacteria.

  涉及假单胞菌HR199（DSM 7063）将香草醛生物转化为原儿茶酸的基因位点vdh、vanA和vanB被鉴定为新型香草醛脱氢酶（vdh）的结构基因和香草酸去甲基酶（vanA和vanB）的两个亚基。这些基因位于EcoRI片段（E230）上，该片段是从cosmid pVK100中的假单胞菌HR199基因组文库中克隆的。vdh基因位于E230的一个亚片段（HE35）上，而vanA和vanB基因则位于E230的另一个亚片段（H110）上。片段HE35和片段H110的一部分的核苷酸序列被确定，揭示了1062、951和1446 bp的开放阅读框，分别代表vanA、vanB和vdh。vdh基因与第四个开放阅读框（ORF2）一起组成一个操纵子，ORF2长735 bp，位于vdh的上游。vanA和vanB的推导氨基酸序列分别与假单胞菌ATCC 19151（F. Brunel和J. Davison，J. Bacteriol. 170:4924-4930，1988）的相应基因产物的氨基酸序列相似度为78.8％和62.1％。vdh基因的推导氨基酸序列与来自不同来源的醛脱氢酶的相似度高达35.3％。ORF2的推导氨基酸序列与烯酰辅酶A水合酶的氨基酸序列相似度高达28.4％。携带在pBluescript SK-中的E230片段的大肠杆菌菌株通过香草酸酯的形式将香草醛转化为原儿茶酸，表明vdh、vanA和vanB在大肠杆菌中具有功能性表达。在pBluescript SK-中克隆的HE35使vdh在大肠杆菌中高表达。由此产生的重组菌株以每毫升培养液最高0.3微摩尔的速率将香草醛转化为香草酸酯。将vanA、vanB和vdh转移到无法利用香草醛或香草酸酯作为碳源的不同假单胞菌菌株和营养生长杆菌中，使这些细菌能够在这些底物上生长。
• Whole-cell bioconversion of vanillin to vanillic acid by Streptomyces viridosporus
  作者：A L Pometto、D L Crawford

  DOI：10.1128/aem.45.5.1582-1585.1983

  日期：1983.5

  cells and was quantitatively oxidized to vanillic acid which accumulated in the growth medium. Vanillic acid was readily recovered from the spent medium by a combination of acid precipitation and ether extraction at greater than or equal to 96% molar yield and upon recrystallization from glacial acetic acid was obtained in greater than or equal to 99% purity.

  利用链霉菌T7A的全细胞，开发了两步分批发酵-香草醛（4-羟基-3-甲氧基苯甲醛）生物转化为香草酸（4-羟基-3-甲氧基苯甲酸）。第一步，在细胞产生芳香醛氧化酶的条件下，使细胞在酵母提取物香兰素培养基中生长。在第二步中，将香草醛与活性细胞孵育，并定量地氧化为在生长培养基中积累的香草酸。通过酸沉淀和醚萃取的组合，以大于或等于96％的摩尔收率容易地从用过的培养基中回收香草酸，并从冰醋酸中重结晶后获得纯度大于或等于99％的乙酸。